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. 2014 Oct;6(4):312-9.
doi: 10.4103/0974-8490.138280.

Antiproliferative effects of Plumbago rosea and its purified constituent plumbagin on SK-MEL 28 melanoma cell lines

Alexander Ronaldo Anuf  1 , Rajesh Ramachandran  2 , Rajaram Krishnasamy  3 , P S Sudhakar Gandhi  3 , Sureshkumar Periyasamy  3
Affiliations

Antiproliferative effects of Plumbago rosea and its purified constituent plumbagin on SK-MEL 28 melanoma cell lines

Alexander Ronaldo Anuf et al. Pharmacognosy Res. 2014 Oct.

Abstract

Background: Plumbago rosea is used in traditional systems of medicine for the preparation of formulations used for treating inflammations, cough, bronchitis, and gastrointestinal disorders, and also in conjunction with cancer chemotherapy. In the present study, the cytotoxic and anti-proliferative effects of plumbagin, and the ethanolic root extract of P. rosea (ETPR) was evaluated on SK-MEL 28 melanoma cell lines and human lymphocytes.

Materials and methods: MTT and apoptotic assays were used for the evaluation of cytotoxic and anti-proliferative effects, respectively. In addition, the effect of Plumbagin and ETPR in down regulation of BCL-2 expression is investigated using RT-PCR analysis.

Results: Both plumbagin and ETPR dose-dependently decreased the cell viability more potently in melanoma cell lines. P. rosea extract demonstrated significant synergy in inhibiting BCL-2 expression than plumbagin. Moreover plumbagin showed more toxicity in human lymphocytes.

Conclusion: Plumbagin has anti-cancer potential, but the side effects limits its use; yet plumbagin, in combination with other ingredients in Plumbago rosea extract, displays significant synergy leading to a stronger anticancer effect with significantly less toxicity.

Keywords: BCL-2; Plumbago rosea; SK-MEL 28; ethanolic root extract of P. rosea; high performance liquid chromatography; plumbagin.

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Conflict of interest statement

Conflict of Interest: None declared

Figures

Figure 1
Figure 1
Qunatification of plumbagin in ethanolic extracts of Plumbago rosea roots using HPLC
Figure 2
Figure 2
Cytotoxic effect of (a) ETPR, (b) Plumbagin on human lymphocytes cells after 48-h treatment. Each data point represents the mean from three independent experiments (mean ± SE)
Figure 3
Figure 3
Antiproliferative effect of (a) ETPR, (b) Plumbagin on SK-MEL 28 cells after 24-h treatment. Data are expressed as a percentage of control (statistics data). Each data point represents the mean of four wells from three independent experiments (mean ± SE)
Figure 4
Figure 4
(i) Colony formation and cell growth in SK-MEL28 melanoma cell lines treated with plumbagin & ETPR, (a) control, (b) 400 μg/mL of ETPR, (c) 10 μg/mL of plumbagin in DMEM containing 10% FBS, (ii) Cell numbers determined by hemocytometer. Values are expressed as mean ± SE from three independent experiments
Figure 5
Figure 5
(i) Fluorescence photomicrographs of SK-MEL 28 melanoma cell lines stained with acridine orange and ethidium bromide, after being treated with ETPR and plumbagin. (a) Control cells with bright red nucleus, (b) cells treated with 10 μg/ml plumbagin, (c) cells treated with 400 μg/ml ETPR. ii) The bar graph shows the percentage of apoptotic cells. The baseline apoptosis in the untreated group was normalized with data from the treated group. The data shown are representative of the combined means from three independent experiments
Figure 6
Figure 6
(i) Levels of RNA transcript of BCL-2 was decreased markedly in 400 μg/ml ETPR-treated cells when compared to 10 μg/ml plumbagintreated cells. GAPDH was used as internal control, (ii) Densitometric analysis of RNA transcript using IMAGE VIEW™ software. Represented data values were obtained from triplicate analysis and expressed as the mean ± SD
See this image and copyright information in PMC

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